Method of making patient specific anti-idiotype antibodies
a technology of anti-idiotype antibodies and patient, applied in the field of humanized tumortargeting molecules in cancer therapeutics, can solve problems such as serious side effects
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example 1
of Cancer in a Subject with Un-Mutated Chronic Lymphocytic Leukemia (CLL)
[0267]A subject presents with multiple symptoms, including fever, fatigue, and enlarged lymph nodes. A clinician non-invasively obtains a blood sample to perform a peripheral blood smear and flow cytometry, ultimately diagnosing the subject with chronic lymphocytic leukemia (CLL). A portion of the blood sample is fractionated to isolate the buffy coat, and deep sequencing of genomic DNA from the CLL leukemia B-cells is performed, thereby determining that the IGHV1-69 allele exhibits greater than 98% sequence homology to the germline IGHV gene. Prognosticating an aggressive disease course, the clinician prescribes a personalized therapy using an antibody that binds to an Fc receptor on an effector cell and the specific gene product of the IGHV1-69 allele sequenced from the subject's CLL B-cells. The subject is pre-medicated with diphenhydramine (50 mg) and acetaminophen (500 mg to 1000 mg) 30 minutes prior to fi...
example 2
of an Autoimmune Disease in a Subject
[0268]A subject is diagnosed with rheumatoid arthritis, a disease characterized by the presence of abnormal anti-self antibodies produced by a single abnormal clonal lineage of B lymphocytes that express the antibodies on their cell surface. A biopsy of the diseased B lymphocytes is obtained, and the sequence of the idiotype, including assessment of microheterogeneity, is determined through selective sequence amplification followed by molecular sequence analysis. The amplicons encoding the idiotype genes are used to develop an anti idiotype therapeutic antibody having one domain with an affinity for the abnormal anti-self antibody produced by diseased B lymphocytes, and a second domain with an affinity for an Fc receptor (e.g., such as those found on effector cells). When administered the subject, the therapeutic antibody induces immune cells to attack and kill the clonal lineage of B lymphocytes producing the abnormal anti-self antibodies, there...
example 3
l Antibody Production Using Hybridoma Technology
[0269]Hybridomas can be used to make the constructs (e.g., antibodies) of the present disclosure, and are generated by immunizing mice with live patient derived tumor cells or membrane extracts made therefrom. The mice are inoculated intraperitoneally with an immunogenic amount of the cells or extract and then boosted with similar amounts of the immunogen. Spleens are collected from the immunized mice, and hybridomas are prepared from the splenocytes and a murine tumor partner using a somatic cell hybridization technique. Tumor cells and splenocytes are fused using the fusogen polyethylene glycol. The fused cells are separated from the fusion medium and grown in a HAT selective growth medium to eliminate unhybridized parent cells. The hybridomas are expanded, and supernatants are assayed for anti-tumor activity by solid-phase enzyme-linked immunosorbent assay (ELISA) using the immunizing agent (tumor cells or membrane extracts made the...
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